论文部分内容阅读
目的 获得阴道毛滴虫Rac1蛋白的cDNA克隆 ,研究其在细胞周期中的调解作用。 方法 提取阴道毛滴虫总RNA ,构建cDNA表达文库 ,随机分离cDNA克隆并测序。用在线生物分析软件NCBIBLAST、ClustalW以及Treeview等程序进行序列分析。 结果 获得一株有 714bp的cDNA克隆。序列分析表明 ,该克隆开放阅读框具 60 0bp ,推测肽链具 2 0 0个氨基酸。该肽链与Rho家族中Rac1鸟苷三磷酸 (GTP)酶同源性最高 (>60 % ) ,并具多种RhoGTP酶的保守基序 ,如GTP结合部位、GTP酶激活蛋白作用基序、GTP分离抑制因子作用基序、鸟嘌呤核苷酸交换因子作用基序等。进化树分析显示该克隆属于Rac亚家族GTP酶 ,与原虫Rac1蛋白最接近。 结论 该克隆属RhoGTP酶的Rac亚家族 ,很可能是阴道毛滴虫的Rac1蛋白。
Objective To obtain the cDNA clone of Trichomonas vaginalis Rac1 protein and study its role in the cell cycle mediation. Methods Total RNA was extracted from Trichomonas vaginalis, cDNA expression library was constructed and cDNA clones were randomly separated and sequenced. Sequence analysis was performed using on-line bioanalysis software NCBIBLAST, ClustalW and Treeview. As a result, a cDNA clone of 714 bp was obtained. Sequence analysis showed that the cloned open reading frame 60 0bp, suggesting that the peptide chain with 200 amino acids. The peptide chain has the highest homology (> 60%) with the Rac1 guanosine triphosphate (GTP) enzyme in the Rho family and has many conserved motifs of RhoGTPase such as GTP binding site, GTPase activating protein action motif, GTP inhibitor of action of the role of motifs, guanine nucleotide exchange factor motifs and so on. Phylogenetic analysis showed that the clone belongs to the Rac subfamily GTPase, which is closest to the protozoal Rac1 protein. Conclusion This clone belongs to the Rac subfamily of RhoGTPases and is likely to be the Rac1 protein of Trichomonas vaginalis.